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Francisellosis of Atlantic Cod (Gadus Morhua L.) ICES IDENTIFICATION LEAFLETS FOR DISEASES AND PARASITES OF FISH AND SHELLFISH Leaflet No. 64 Francisellosis of Atlantic cod (Gadus morhua L.) Anders Alfjorden and Neil Ruane International Council for the Exploration of the Sea Conseil International pour l’Exploration de la Mer H.C. Andersens Boulevard 44–46 DK-1553 Copenhagen V Denmark Telephone (+45) 33 38 67 00 Telefax (+45) 33 93 42 15 www.ices.dk [email protected] Recommended format for purposes of citation: Alfjorden, A., and Ruane, N. 2015. Francisellosis of Atlantic cod (Gadus morhua L.). ICES Identification Leaflets for Diseases and Parasites of Fish and Shellfish. Leaflet No. 64. 5 pp. Series Editor: Stephen Feist. Prepared under the auspices of the ICES Working Group on Pathology and Diseases of Marine Organisms. The material in this report may be reused for non-commercial purposes using the recommended citation. ICES may only grant usage rights of information, data, images, graphs, etc. of which it has ownership. For other third-party material cited in this report, you must contact the original copyright holder for permission. For citation of datasets or use of data to be included in other databases, please refer to the latest ICES data policy on the ICES website. All extracts must be acknowledged. For other reproduction requests please contact the General Secretary. ISBN 978-87-7482-173-1 ISSN 0109–2510 © 2015 International Council for the Exploration of the Sea Leaflet No. 64 | 1 Francisellosis of Atlantic cod (Gadus morhua L.) Anders Alfjorden and Neil Ruane Susceptible species Francisellosis, caused by infection with Francisella noatunensis, primarily affects Atlantic cod (Gadus morhua) and was first described in Norway in 2004 (Nylund et al., 2006) and 2005 (Olsen et al., 2006). The disease has also been reported affecting Atlantic salmon (Salmo salar) in Chile (Birkbeck et al., 2007). Disease name Francisellosis. Aetiological agent F. noatunensis is recognized as the causative agent of francisellosis in Atlantic cod (Ottem et al., 2009). Bacteria belonging to the genus Francisella are Gram-negative, non- motile, aerobic, facultative intracellular organisms belonging to the γ-proteobacteria. Francisella sp. are generally biochemically unreactive and the number of phenotypical tests useful for differentiation of the species are few (Colquhoun and Duodo, 2011). Fish pathogenic Francisella sp. can be distinguished by their requirement for cysteine in culture media and their inability to grow at temperatures above 35°C (Ottem et al., 2007). Geographical distribution The disease has been reported primarily from farmed Atlantic cod in Norway (Nylund et al., 2006; Olsen et al., 2006) and from wild-caught cod from the South coast of Ireland and subsequently reared in a research facility (Ruane et al., 2015). There is also evidence that the bacterium is present in wild Atlantic cod along the west coasts of Sweden and Norway, with a reported prevalence of 6.8% off southwest Norway (Alfjorden et al., 2006; Ottem et al., 2008; Colquhoun and Duodo, 2011). A visceral granulomatous disease in Atlantic cod in the North Sea in the 1980s was also reported to be due to F. noatunensis (Zerihun et al., 2011). The disease has also been reported causing mortalities in freshwater farmed Atlantic salmon in Chile (Birkbeck et al., 2007). Associated environmental conditions There is little information available on the environmental conditions associated with clinical Francisellosis. Clinical disease has been reported in Atlantic cod at water temperatures above 14oC (Olsen et al., 2006; Ruane et al., 2015) and Ottem et al., (2008) suggested that the differences in disease prevalence in cod farms in the north and south of Norway could be temperature related. Significance Francisellosis is a significant health problem for Atlantic cod aquaculture, particularly in Norway which has seen a decline in the number of registered farms from 240 in 2007 to 84 in 2012 (Hjeltnes, 2014). As the disease is chronic in nature and highly infectious, the prevalence of infected fish in a population can be high. 2 | Francisellosis of Atlantic cod (Gadus morhua L.) Gross clinical signs Francisellosis is principally a chronic systemic granulomatous inflammatory disease with varying degrees of mortality. The gross signs observed among most host species are similar and include disseminated white nodules of various sizes in liver, spleen and kidney although most tissues and organs may be affected. Splenomegaly and sero- sanguinous ascites has been observed in cod. Microscopic lesions include widespread chronic granulomatous inflammation in all organs, associated with variable numbers of Gram-negative bacteria (Birkbeck et al., 2011). Control measures and legislation Francisellosis is not reportable to the OIE but is a List 3 disease in Norway. There is currently no licensed vaccine available for the disease although research on a trial vaccine for francisellosis caused by F. asiatica in tilapia, Oreochromis niloticus has shown promise (Soto et al., 2011). Due to the intracellular nature of the bacterium, antibiotic treatment has proven difficult and there are no reports of successful antibiotic treatments of francisellosis in Atlantic cod. Diagnostic methods The gold standard for diagnosis of francisellosis is culture of the bacterium combined with macroscopic and histological observations of affected tissues (Colquhoun and Duodo, 2011). F. noatunensis does not grow on routine, general-purpose agar plates and is also easily outcompeted by other bacteria. Several agars selective for Francisella sp. have been published (Birkbeck et al., 2011), the most common being cysteine heart agar supplemented with antibiotics. Immunohistological methods have also been used to diagnose this disease (Jansson et al., 2007). Diagnosis using molecular methods is also common for francisellosis including sequencing of the 16S rRNA gene (Nylund et al., 2006; Olsen et al., 2006), real-time PCR (Ottem et al., 2008) and in situ hybridization (Colquhoun and Duodu, 2011). Key References Alfjorden, A., Jansson E., Johansson K.-E. 2006. A systemic granulomatous inflammatory disease in wild Atlantic cod, Gadus morhua associated with a bacterium of the genus Francisella. DIPNET, Disease interaction and pathogen exchange between farmed and wild aquatic animal population, A European network, Newsletter 44. Birkbeck, T. H., Bordevik, M., Frøystad, M. K., Baklien, Å. 2007. Identification of Francisella sp. from Atlantic salmon, Salmo salar L., in Chile. Journal of Fish Diseases, 30: 505–507. Birkbeck, T. H., Feist, S. W., Verner-Jeffreys, D. W. 2011. Francisella infections in fish and shellfish. Journal of Fish Diseases, 34: 173–187. Colquhoun, D. J., and Duodo, S. 2011. Francisella infections in farmed and wild aquatic organisms. Veterinary Research, 42: 47. Hjeltnes, B. 2014. The Health Situation in Norwegian Aquaculture 2013. Norwegian Veterinary Institute, Oslo, 42pp. Jansson, E., Alfjorden A., Kjellberg E., Mattsson R., Ottander L., Hellström, A., 2007. Diagnosis of Francisella sp. infection in wild Atlantic cod, Gadhus morhua, by immunohistochemistry. Proceedings from E.A.F.P. 13th International Conference, Grado, Italy. Leaflet No. 64 | 3 Nylund, A., Ottem, K. F., Watanabe, K., Karlsbakk, E., and Krossøy, B. 2006. Francisella sp. (Family Francisellaceae) causing mortality in Norwegian cod (Gadus morhua) farming. Archives of Microbiology, 185: 383–392. Olsen, A. B., Mikalsen, J., Rode, M., Alfjorden, A., Hoel, E., Straum-Lie, K., Haldorsen, R., and Colquhoun, D. J. 2006. A novel systemic granulomatous inflammatory disease in farmed Atlantic cod, Gadus morhua L., associated with a bacterium belonging to the genus Francisella. Journal of Fish Diseases, 29: 307–311. Ottem, K. F., Nylund, A., Karlsbakk, E., Friis-Møller, A., and Kamaishi, T. 2009. Elevation of Francisella philomiragia subsp. noatunensis Mikalsen et al. (2007) to Francisella noatunensis comb. nov. [syn. Francisella piscicida Ottem et al. (2008) syn. nov.] and characterization of Francisella noatunensis subsp. orientalis subsp. nov., two important fish pathogens. Journal of Applied Microbiology, 106: 1231–1243. Ottem, K. F., Nylund, A., Isaksen, T. E., Karlsbakk, E., and Bergh, Ø. 2008. Occurrence of Francisella piscicida in farmed and wild Atlantic cod, Gadus morhua L., in Norway. Journal of Fish Diseases, 31: 525–534. Ottem, K. F., Nylund, A., Karlsbakk, E., Friis-Møller, A., and Krossøy, B. 2007. Characterization of Francisella sp., GM2212, the first Francisella isolate from marine fish, Atlantic cod (Gadus morhua). Archives of Microbiology, 187: 343–350. Ruane, N. M., Bolton-Warberg, M., Rodger, H. D., Colquhoun, D. J., Geary, M., McCleary, S. J., O Halloran, K., Maher, K., O Keeffe, D., Mirimin, L., Henshilwood, K., Geoghegan, F., and Fitzgerald, R. D. 2015. An outbreak of francisellosis in wild- caught Celtic Sea Atlantic cod, Gadus morhua L., juveniles reared in captivity. Journal of Fish Diseases 38: 97–102. Soto, E., Wiles, J., Elzer, P., Macaluso, K., Hawke, J. P. 2011. Attenuated Francisella asiatica iglC mutant induces protective immunity to francisellosis in tilapia. Vaccine, 29: 593–598. Zerihun, M. A., Feist, S. W., Bucke, D., Olsen, A. B., Tandstad, N. M., and Colquhoun, D. J. 2011. Francisella noatunensis subsp. noatunensis is the aetiological agent of visceral granulomatosis in wild Atlantic cod Gadus morhua. Diseases of Aquatic Organisms, 95: 65–71. 4 | Francisellosis of Atlantic cod (Gadus morhua L.) Figure 1. Gross clinical appearance of Atlantic cod, Gadus
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